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Functional analysis of the Candida albicans ECE1 Promoter

The formation of hyphae is a key virulence attribute of Candida albicans as they are required for adhesion to and invasion of host cells, and ultimately deep-tissue dissemination. Hyphae also secrete the peptide toxin candidalysin, which is crucial for destruction of host cell membranes. The peptide...

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Autores principales: Garbe, Enrico, Thielemann, Nadja, Hohner, Sina, Kumar, Animesh, Vylkova, Slavena, Kurzai, Oliver, Martin, Ronny
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10100963/
https://www.ncbi.nlm.nih.gov/pubmed/36786567
http://dx.doi.org/10.1128/spectrum.00253-23
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author Garbe, Enrico
Thielemann, Nadja
Hohner, Sina
Kumar, Animesh
Vylkova, Slavena
Kurzai, Oliver
Martin, Ronny
author_facet Garbe, Enrico
Thielemann, Nadja
Hohner, Sina
Kumar, Animesh
Vylkova, Slavena
Kurzai, Oliver
Martin, Ronny
author_sort Garbe, Enrico
collection PubMed
description The formation of hyphae is a key virulence attribute of Candida albicans as they are required for adhesion to and invasion of host cells, and ultimately deep-tissue dissemination. Hyphae also secrete the peptide toxin candidalysin, which is crucial for destruction of host cell membranes. The peptide is derived from a precursor protein encoded by the gene ECE1 which is strongly induced during hyphal growth. Previous studies revealed a very complex regulation of this gene involving several transcription factors. However, the promoter of the gene is still not characterized. Here, we present a functional analysis of the intergenic region upstream of the ECE1 gene. Rapid amplification of cDNA ends (RACE)-PCR was performed to identify the 5′ untranslated region, which has a size of 49 bp regardless of the hyphae-inducing condition. By using green fluorescent protein (GFP) reporter constructs we further defined a minimal promoter length of 1,500 bp which was verified by RT-qPCR. Finally, we identified the TATA element required for the expression of the gene. It is located 106 to 109 bp upstream of the ECE1 start codon. Our results illustrate that despite a very short 5′ UTR, a relatively long promoter is required to secure ECE1 transcription, indicating a complex regulatory machinery tightly controlling the expression of the gene. IMPORTANCE In recent years it was shown that secretion of the toxic peptide candidalysin from hyphae of the major human fungal pathogen Candida albicans contributes heavily to its virulence. The peptide is derived from a precursor protein which is encoded by the ECE1 gene whose transcription is known to be closely associated with formation of hyphae. Here, we used a GFP reporter system to determine the length of the ECE1 promoter and were able to show that it has a minimal size of 1,500 bp. Surprisingly, the gene has a very short 5′ UTR of only 49 bp. In accordance with this, the TATA element required for transcription is located 106 to 109 bp upstream of the start codon. This indicates that ECE1 expression is controlled by a very long promoter allowing a complex network of transcription factors to contribute to the gene’s regulation.
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spelling pubmed-101009632023-04-14 Functional analysis of the Candida albicans ECE1 Promoter Garbe, Enrico Thielemann, Nadja Hohner, Sina Kumar, Animesh Vylkova, Slavena Kurzai, Oliver Martin, Ronny Microbiol Spectr Research Article The formation of hyphae is a key virulence attribute of Candida albicans as they are required for adhesion to and invasion of host cells, and ultimately deep-tissue dissemination. Hyphae also secrete the peptide toxin candidalysin, which is crucial for destruction of host cell membranes. The peptide is derived from a precursor protein encoded by the gene ECE1 which is strongly induced during hyphal growth. Previous studies revealed a very complex regulation of this gene involving several transcription factors. However, the promoter of the gene is still not characterized. Here, we present a functional analysis of the intergenic region upstream of the ECE1 gene. Rapid amplification of cDNA ends (RACE)-PCR was performed to identify the 5′ untranslated region, which has a size of 49 bp regardless of the hyphae-inducing condition. By using green fluorescent protein (GFP) reporter constructs we further defined a minimal promoter length of 1,500 bp which was verified by RT-qPCR. Finally, we identified the TATA element required for the expression of the gene. It is located 106 to 109 bp upstream of the ECE1 start codon. Our results illustrate that despite a very short 5′ UTR, a relatively long promoter is required to secure ECE1 transcription, indicating a complex regulatory machinery tightly controlling the expression of the gene. IMPORTANCE In recent years it was shown that secretion of the toxic peptide candidalysin from hyphae of the major human fungal pathogen Candida albicans contributes heavily to its virulence. The peptide is derived from a precursor protein which is encoded by the ECE1 gene whose transcription is known to be closely associated with formation of hyphae. Here, we used a GFP reporter system to determine the length of the ECE1 promoter and were able to show that it has a minimal size of 1,500 bp. Surprisingly, the gene has a very short 5′ UTR of only 49 bp. In accordance with this, the TATA element required for transcription is located 106 to 109 bp upstream of the start codon. This indicates that ECE1 expression is controlled by a very long promoter allowing a complex network of transcription factors to contribute to the gene’s regulation. American Society for Microbiology 2023-02-14 /pmc/articles/PMC10100963/ /pubmed/36786567 http://dx.doi.org/10.1128/spectrum.00253-23 Text en Copyright © 2023 Garbe et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Garbe, Enrico
Thielemann, Nadja
Hohner, Sina
Kumar, Animesh
Vylkova, Slavena
Kurzai, Oliver
Martin, Ronny
Functional analysis of the Candida albicans ECE1 Promoter
title Functional analysis of the Candida albicans ECE1 Promoter
title_full Functional analysis of the Candida albicans ECE1 Promoter
title_fullStr Functional analysis of the Candida albicans ECE1 Promoter
title_full_unstemmed Functional analysis of the Candida albicans ECE1 Promoter
title_short Functional analysis of the Candida albicans ECE1 Promoter
title_sort functional analysis of the candida albicans ece1 promoter
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10100963/
https://www.ncbi.nlm.nih.gov/pubmed/36786567
http://dx.doi.org/10.1128/spectrum.00253-23
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